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Abstract Embryo implantation failures are a major challenge in reproductive medicine, but the underlying mechanism remains poorly understood. Successful implantation requires dynamic remodeling of the endometrium through integrated proliferation and differentiation of endometrial cells including luminal epithelial, glandular epithelial, and stromal cells. Conversely, their disruption causes infertility. Spatiotemporal control of transcription is required for these processes; however, the underlying epigenetic regulation is largely unknown. In this study, we examined expression data from the human endometrium during implantation and discovered that expression of the histone lysine methyltransferase KMT2D was significantly suppressed in patients with recurrent implantation failure. Further study revealed that uterine deletion of Kmt2d in mice caused infertility due to implantation failure. Morphological analysis discovered a reduction in the number of uterine glands and aberrant differentiation of the luminal and glandular epithelium into stratified phenotypes in Kmt2d knockout uteri. Administration of leukemia inhibitory factor protein, which is expressed in uterine glands and is essential for implantation, did not rescue implantation failure in Kmt2d knockout mice, suggesting that infertility was not solely due to uterine gland dysfunction. RNA sequencing analysis revealed that Kmt2d knockout uteri displayed suppressed expression of genes involved in ion homeostasis, which may affect the uterine luminal morphology. Our study suggests that KMT2D plays an essential role in facilitating successful embryo implantation by regulating the coordinated differentiation of endometrial cells, providing valuable insights into unexplained implantation failures in women.

BackgroundThe global obesity epidemic is a significant public health issue, often leading to metabolic disorders such as diabetes and cardiovascular diseases. Collagen peptides (CP) and their bioactive component, Prolyl-hydroxyproline (Pro-Hyp), have shown potential in reducing adipocyte size, with unclear mechanisms concerning brown adipocyte differentiation.MethodsWe investigated the effects of Pro-Hyp on the differentiation of brown adipocytes in C3H10T1/2 mesenchymal stem cells, focusing on its impact on adipocyte size, gene expression related to brown fat function, and mitochondrial activity.ResultsPro-Hyp treatment decreased adipocyte size and upregulated brown fat-specific genes, including C/EBPα, PGC-1α, and UCP-1. Remarkably, it did not alter PPARγ expression. Pro-Hyp also elevated mitochondrial activity, suggesting enhanced brown adipocyte functionality. A Pro-Hyp responsive element was identified in the PGC-1α gene promoter, which facilitated the binding of the Foxg1 transcription factor, indicating a novel regulatory mechanism.ConclusionPro-Hyp promotes brown adipocyte differentiation, potentially offering a therapeutic strategy for obesity management. This study provides a molecular basis for the anti-obesity effects of CP, although further in vivo studies are needed to confirm these findings and to investigate the potential impact on beige adipocyte differentiation.

The production of cell-type– and age-specific genetically modified mice is a powerful approach for unraveling unknown gene functions. Here, we present a simple and timesaving method that enables adeno-associated virus (AAV)–mediated cell-type– and age-specific recombination in floxed mice. To achieve astrocyte-specific recombination in floxed Ai14 reporter mice, we intravenously injected blood-brain barrier–penetrating AAV-PHP.eB vectors expressing Cre recombinase (Cre) using the astrocyte-specific mouse glial fibrillary acidic protein (mGfaABC1D) promoter. However, we observed nonspecific neuron-predominant transduction despite the use of an astrocyte-specific promoter. We speculated that subtle but continuous Cre expression in nonastrocytic cells triggers recombination, and that excess production of Cre in astrocytes inhibits recombination by forming Cre-DNA aggregates. Here, we resolved this paradoxical event by dividing a single AAV into two mGfaABC1D-promoter-driven AAV vectors, one expressing codon-optimized flippase (FlpO) and another expressing flippase recognition target–flanked rapidly degrading Cre (dCre), together with switching the neuron-tropic PHP.eB capsid to astrocyte-tropic AAV-F. Moreover, we found that the FlpO-dCre system with a target cell-tropic capsid can also function in neuron-targeting recombination in floxed mice.

Endoscopic retrograde cholangiopancreatography (ERCP) in patients with Roux-en-Y gastric bypass anatomy is a well-documented challenge. Traditionally, this problem has been overcome with adjunctive techniques, such as device-assisted ERCP, including double-balloon or single-balloon enteroscopy and laparoscopy-assisted transgastric ERCP. Endoscopic ultrasound-directed transgastric ERCP (EDGE) is a novel technique that enables access to the ampulla using a duodenoscope without surgical intervention and has shown high clinical and technical success rates in recent studies. However, this approach is technically demanding, necessitating a thorough understanding of the gastrointestinal anatomy as well as high operator experience. In this review, we provide a technical overview of EDGE in parallel with our personal experience at our center and propose a simple algorithm to select patients for its appropriate application. In conjunction, the outcomes of EDGE compared with those of device-assisted and laparoscopy-assisted transgastric ERCP will be discussed.

In this work, Yoshinaga et al. use functional CRISPR screens and in vivo analyses to show that an RNA methyltransferase METTL16 and MTR4-nuclear exosome play an essential role in maintaining erythropoiesis and regulating DNA repair in erythroblasts.

Strabismus, a neuro-ophthalmological condition characterized by misalignment of the eyes, is a common ophthalmic disorder affecting both children and adults. In our previous study, we identified the microsomal glutathione S-transferase 2 (MGST2) gene as one of the potential candidates for comitant strabismus susceptibility in a Japanese population. The MGST2 gene belongs to the membrane-associated protein involved in the generation of pro-inflammatory mediators, and it is also found in the protection against oxidative stress by decreasing the reactivity of oxidized lipids. To look for the roles of the MGST2 gene in the development, eye alignment, and overall morphology of the eye as the possible background of strabismus, MGST2 gene knockout (KO) mice were generated by CRISPR/Cas9-mediated gene editing with guide RNAs targeting the MGST2 exon 2. The ocular morphology of the KO mice was analyzed through high-resolution images obtained by a magnetic resonance imaging (MRI) machine for small animals. The morphometric analyses showed that the height, width, and volume of the eyeballs in MGST2 KO homozygous mice were significantly greater than those of wild-type mice, indicating that the eyes of MGST2 KO homozygous mice were significantly enlarged. There were no significant differences in the axis length and axis angle. These morphological changes may potentially contribute to the development of a subgroup of strabismus.

Abstract Objectives This study aimed to objectively evaluate the water‐jet‐functioned electrosurgical knife injection performances in a desktop experiment. Methods Five types of water‐jet‐functioned electrosurgical knives, including two injection styles of sheath‐type (A: DualKnife J, KD‐655L; B: FlushKnife, DK2620‐J‐B20S; C: Splash M‐Knife, DN‐D2718B; D: ISSEN, SN1650‐20) and tip‐type (E: ORISE ProKnife, M00519361) were evaluated. These knives were compared with an injection needle (Control: SuperGrip 25G) as a control. The injection speed under constant pressure and the injection efficiency for each knife against prepared porcine stomach mucosa were evaluated. The additional clear gel injections using an injection needle were observed using an indigo blue‐colored gel to evaluate the difference between the locations of water‐jet holes. Results Four types of knives, except for A, showed significantly higher water‐jet speeds (A: 0.79 ± 0.03 g/20 s, B: 2.56 ± 0.05 g/20 s, C: 3.09 ± 0.06 g/20 s, D: 2.86 ± 0.05 g/20 s, and E: 1.79 ± 0.03 g/20 s) compared to that of the control (1.21 ± 0.03 g/20 s). Meanwhile, significantly higher efficacy of injection was found in the tip‐type water‐jet function knife, second to the injection needle (Control: 37.2% ± 35.5%, A: 20.9% ± 20.2%, B: 1.1% ± 2.2%, C: 6.2% ± 12.6%, D: 12.5% ± 15.6%, and E: 33.3% ± 32.2%). An additional injection experiment revealed that the injection with a piercing tip into the gel could achieve sufficient additional injection inside the stacked clear gel. Conclusions The tip‐type water‐jet function electrosurgical knife is preferable for effective submucosal injection during endoscopic treatments.

In the maintenance of cable structures, such as cable-stayed bridges, cable safety is assessed based on the cable tension. In Japan, the cable tension is generally estimated from the cable’s natural frequencies using the higher-order vibration method. In recent years, dampers have been installed onto cables to suppress aerodynamic vibrations. Because the damper changes the cable’s natural frequencies, the damper is removed to measure the natural frequencies and estimate the cable tension without a damper, and the damper is then reinstalled. To avoid damper removal and reinstallation, the authors previously proposed Method 2F for estimating the tension of a cable with a damper from the natural frequencies without removing the damper. Because the tension estimation error of the higher-order vibration method for a cable without a damper has been reported as 5%, the authors set the target tension estimation error within 5%. However, the tension estimation error of Method 2F exceeded 5% in the experimental verification. Furthermore, although Method 2F estimates the tension and bending stiffness of the cable and the damper parameters simultaneously from the natural frequencies, the accuracy of the bending stiffness and damper parameters is unsatisfactory. In this paper, the new Method 2FM is proposed to estimate the tension and bending stiffness of the cable and damper parameters using the natural frequencies and two-point mode shapes. With the addition of mode shapes, Method 2FM attempts to improve the accuracy of estimating the tension and other parameters. The validity of Method 2FM was confirmed by numerical simulations and experiments. The numerical verification confirmed that Method 2FM is superior to Method 2F in estimating the cable tension and damper parameters. The experimental verification confirmed that the tension estimation accuracy of Method 2FM is higher than that of Method 2F, and the estimation error is lower than 5%. However, the damper parameters estimated by Method 2FM are different to the design values. The reason for this is the modeling error of the damper, as found by conducting an element test on the damper.

In the maintenance of cable structures, such as cable-stayed bridges, it is necessary to estimate the tension acting on the cables. In current Japanese practice, the cable tension is estimated from the cable’s natural frequency using vibration-based methods. However, in recent years, dampers have been installed onto the cables to suppress aerodynamic vibrations. Because the damper changes the cable’s natural frequencies, the methods used for cables without dampers are not appropriate for cables with dampers. With this background, the authors previously proposed a method (Method 0F) for estimating the tension of a cable with a damper from the natural frequencies and their modal order. Method 0F partially ignores the imaginary part of the complex natural frequencies to simplify the problem. This study proposes a new method (Method 1F) that does not ignore the imaginary part of a complex natural frequency. Method 1F still needs both the natural frequencies and their modal order to be input. If the modal order is not correctly specified, the accuracy deteriorates. Therefore, a new method (Method 2F) that only requires the natural frequencies is also proposed. The validity of the proposed methods was confirmed by numerical simulations and experiments. The numerical and experimental verifications confirmed that the new methods were superior compared with previous methods, and that Method 2F has the highest effectiveness.

In the maintenance of cable structures, such as cable-stayed bridges and extra-dosed bridges, it is necessary to estimate the tension acting on the cables. The safety of a cable is confirmed by checking whether the tension acting on the cable is within the allowable value. In current Japanese practice, the tension of a cable is estimated using the vibration method or the higher-order vibration method, which considers the natural frequencies of the cable. However, in recent years, the aerodynamic vibration of cables caused by wind has become a problem owing to the recent increase in the cable length and low damping performance of the cable itself. To suppress the aerodynamic vibration of cables, dampers are installed onto the cables. Because the damper changes the cable’s natural frequencies, the vibration method and higher-order vibration method are inappropriate for measuring the tension of a cable with a damper. In this paper, a new tension estimation method for a cable with a damper is proposed. To model a cable with a tensioned Bernoulli-Euler beam, theoretical equations for estimating the natural frequencies were derived. The proposed method inversely estimates the tension and bending stiffness of the cable and damper parameters, simultaneously, from the natural frequencies. The validity of the proposed method was confirmed by conducting numerical simulations and experiments. In the numerical verification, the performance of the proposed method was investigated using 80 numerical models. In the experimental verification, the estimation accuracy of the proposed method was investigated by considering 16 test cases. Thus, it was confirmed that the tension estimation accuracy was high, whereas the bending stiffness and damper parameter estimation accuracy was unsatisfactory. The tension estimation error was within 10% in all experimental cases, and within 5% if two test cases are excluded. The results obtained by the numerical and experimental verifications confirmed the effectiveness of the proposed method in tension estimation.

In this study, we examined the role of the eastern bent-winged bat (Miniopterus fuliginosus) in the dispersion of bat adenovirus and bat alphacoronavirus in east Asia, considering their gene flows and divergence times (based on deep-sequencing data), using bat fecal guano samples. Bats in China moved to Jeju Island and/or Taiwan in the last 20,000 years via the Korean Peninsula and/or Japan. The phylogenies of host mitochondrial D-loop DNA was not significantly congruent with those of bat adenovirus (m2XY = 0.07, p = 0.08), and bat alphacoronavirus (m2XY = 0.48, p = 0.20). We estimate that the first divergence time of bats carrying bat adenovirus in five caves studied (designated as K1, K2, JJ, N2, and F3) occurred approximately 3.17 million years ago. In contrast, the first divergence time of bat adenovirus among bats in the 5 caves was estimated to be approximately 224.32 years ago. The first divergence time of bats in caves CH, JJ, WY, N2, F1, F2, and F3 harboring bat alphacoronavirus was estimated to be 1.59 million years ago. The first divergence time of bat alphacoronavirus among the 7 caves was estimated to be approximately 2,596.92 years ago. The origin of bat adenovirus remains unclear, whereas our findings suggest that bat alphacoronavirus originated in Japan. Surprisingly, bat adenovirus and bat alphacoronavirus appeared to diverge substantially over the last 100 years, even though our gene-flow data indicate that the eastern bent-winged bat serves as an important natural reservoir of both viruses.

The generation of conditional knockout mice using the Cre-loxP system is advantageous for the functional analysis of genes. Flanked by two loxP sites (floxed) mice can be directly obtained from fertilized eggs by the CRISPR/Cas9 genome editing system. We previously reported that sequential knock-in (KI) of each loxP site by electroporation (EP) at the 1- and 2-cell embryonic stages increases the number of mice with floxed alleles compared with simultaneous KI. However, EP at the 2-cell stage frequently induced blastomere fusion. These fused embryos cannot develop to term because they are tetraploidized. In this study, we examined the following three conditions to inhibit blastomere fusion by EP at the 2-cell stage: (1) hypertonic treatment, (2) Calcium (Ca2+)-free treatment, and (3) actin polymerization inhibition. Hypertonic treatment of 2-cell stage embryos prevented blastomere fusion and facilitated blastocyst development; however, KI efficiency was decreased. Ca2+-free treatment and actin polymerization inhibition by cytochalasin B (CB) reduced fusion rate, and did not have negative effects on development and KI efficiency. These results suggest that Ca2+-free and CB treatment at the 2-cell stage is effective to generate floxed mice in combination with a sequential EP method.

Background. Vonoprazan affords more clinical benefits than proton pump inhibitors (PPIs) during the healing of gastroduodenal ulcers. However, it remains controversial whether vonoprazan is more effective than PPIs when used to heal artificial ulcers arising after endoscopic submucosal dissection (ESD). Aim. This study investigated the effects of vonoprazan compared with esomeprazole on the healing of post-ESD artificial ulcers. Methods. Sixty patients who underwent gastric ESD between May 2015 and May 2017 were randomized to treatment with vonoprazan (V group) or esomeprazole (E group) for 8 weeks. Upper endoscopy was performed at 4 and 8 weeks after ESD, and drug effects were estimated based on the ulcer healing rates and shrinkage rates. Results. Fifty-three patients were analyzed. The respective 4- and 8-week ulcer healing rates did not differ significantly between V and E groups (8.0 versus 11.5%, P=0.669; 88.9 versus 84.6%, P=0.420). Similarly, the respective 4- and 8-week ulcer shrinkage rates did not differ significantly between V and E groups (96.8 versus 97.5%, P=0.656; 100 versus 100%, P=0.257). Conclusion. The healing of artificial ulcers after ESD did not differ using vonoprazan or esomeprazole. Both vonoprazan and esomeprazole were effective when used to promote artificial ulcer healing after ESD.

Background and study aims Endoscopic submucosal dissection (ESD) is used to treat superficial colorectal tumors. Previous studies have reported the efficacy of ESD for treating residual or local recurrent colorectal tumors. This study sought to evaluate the efficacy of ESD in treating these lesions and to assess factors that prevent successful ESD. Methods This retrospective study assessed 25 cases of residual or local recurrent lesions that were previously treated using EMR (18 lesions), TEM (5 lesions), ESD (1 lesion) or surgery (1 lesion), and 459 primary lesions treated using ESD between April 2008 and September 2015. Clinicopathological characteristics, treatment outcome and adverse events were compared between groups with or without scar tissue. Factors related to perforation and a prolonged treatment time, which indicate the likelihood of technical difficulties, were identified using multiple logistic regression analysis. Results In residual or local recurrent lesions groups, patients experienced more perforations (32 % vs 4 %, P

We performed post-marketing surveillance to evaluate the safety and efficacy of cell-free and concentrated ascites reinfusion therapy (CART). In total, 356 CART sessions in 147 patients at 22 centers were performed. The most common primary disease was cancer (128 cases, 300 sessions). Mean amount of ascites collected was 3.7 L, and mean concentration ratio was 9.2. Mean amount of reinfused protein was 67.8 g (recovery rate, 72.0%). Performance status, dietary intake, urine volume, body weight and abdominal circumference were significantly improved after CART. Body temperature increased significantly, by 0.3°C on average. Concomitant steroids and/or NSAIDs use before reinfusion was significantly and negatively associated with increases in body temperature. Most adverse events were fever and chills. This study examined a large number of patients compared with previous studies, and showed that CART is an effective and relatively safe treatment for refractory ascites, such as malignant ascites.